Inhibitors of p38

ABSTRACT

The present invention relates to inhibitors of p38, a mammalian protein kinase involved cell proliferation, cell death and response to extracellular stimuli. The invention also relates to methods for producing these inhibitors. The invention also provides pharmaceutical compositions comprising the inhibitors of the invention and methods of utilizing those compositions in the treatment and prevention of various disorders.

TECHNICAL FIELD OF INVENTION

The present invention relates to inhibitors of p38, a mammalian proteinkinase involved in cell proliferation, cell death and response toextracellular stimuli. The invention also relates to methods forproducing these inhibitors. The invention also provides pharmaceuticalcompositions comprising the inhibitors of the invention and methods ofutilizing those compositions in the treatment and prevention of variousdisorders.

BACKGROUND OF THE INVENTION

Protein kinases are involved in various cellular responses toextracellular signals. Recently, a family of mitogen-activated proteinkinases (MAPK) has been discovered. Members of this family are Ser/Thrkinases that activate their substrates by phosphorylation [B. Stein etal., Ann. Rep. Med. Chem., 31, pp. 289-98 (1996)]. MAPKs are themselvesactivated by a variety of signals including growth factors, cytokines,UV radiation, and stress-inducing agents.

One particularly interesting MAPK is p38. p38, also known as cytokinesuppressive anti-inflammatory drug binding protein (CSBP) and RK, wasisolated from murine pre-B cells that were transfected with thelipopolysaccharide (LPS) receptor, CD14, and induced with LPS. p38 hassince been isolated and sequenced, as has the cDNA encoding it in humansand mouse. Activation of p38 has been observed in cells stimulated bystress, such as treatment of lipopolysaccharides (LPS), UV, anisomycin,or osmotic shock, and by cytokines, such as IL-1 and TNF.

Inhibition of p38 kinase leads to a blockade on the production of bothIL-1 and TNF. IL-1 and TNF stimulate the production of otherproinflammatory cytokines such as IL-6 and IL-8 and have been implicatedin acute and chronic inflammatory diseases and in post-menopausalosteoporosis [R. B. Kimble et al., Endocrinol., 136, pp. 3054-61(1995)].

Based upon this finding, it is believed that p38, along with otherMAPKs, have a role in mediating cellular response to inflammatorystimuli, such as leukocyte accumulation, macrophage/monocyte activation,tissue resorption, fever, acute phase responses and neutrophilia. Inaddition, MAPKs, such as p38, have been implicated in cancer,thrombin-induced platelet aggregation, immunodeficiency disorders,autoimmune diseases, cell death, allergies, osteoporosis andneurodegenerative disorders. Inhibitors of p38 have also been implicatedin the area of pain management through inhibition of prostaglandinendoperoxide synthase-2 induction. Other diseases associated with Il-1,IL-6, IL-8 or TNF overproduction are set forth in WO 96/21654.

Others have already begun trying to develop drugs that specificallyinhibit MAPKs. For example, PCT publication WO 95/31451 describespyrazole compounds that inhibit MAPKs, and, in particular, p38. However,the efficacy of these inhibitors in vivo is still being investigated.

Accordingly, there is still a great need to develop other potentinhibitors of p38, including p38-specific inhibitors, that are useful intreating various conditions associated with p38 activation.

SUMMARY OF THE INVENTION

The present invention addresses this problem by providing compounds thatdemonstrate strong inhibition of p38.

These compounds have the general formula:

wherein each of Q₁ and Q₂ are independently selected from a phenyl or5-6 membered aromatic heterocyclic ring system, or a 8-10 memberedbicyclic ring system comprising aromatic carbocyclic rings, aromaticheterocyclic rings or a combination of an aromatic carbocyclic ring andan aromatic heterocyclic ring.

A heterocyclic ring system or a heterocyclic ring contains 1 to 4heteroatoms, which are independently selected from N, O, S, SO and SO₂.

The rings that make up Q₁ are substituted with 1 to 4 substituents, eachof which is independently selected from halo; C₁-C₃ alkyl optionallysubstituted with NR′₂, OR′, CO₂R′ or CONR′₂; O—(C₁-C₃)-alkyl optionallysubstituted with NR′₂, OR′, CO₂R′ or CONR′₂; NR′₂; OCF₃; CF₃; NO₂;CO₂R′; CONR′; SR′; S(O₂)N(R′)₂; SCF₃; CN; N(R′)C(O)R⁴; N(R′)C(O)OR⁴;N(R′)C(O)C(O)R⁴; N(R′)S(O₂)R⁴; N(R′)R⁴; N(R⁴)₂; OR⁴; OC(O)R⁴; OP(O)₃H₂;or N═C—N(R′)₂.

The rings that make UP Q₂ are optionally substituted with up to 4substituents, each of which is independently selected from halo; C₁-C₃straight or branched alkyl optionally substituted with NR′₂, OR′, CO₂R′,S(O₂)N(R′)₂, N═C—N(R′)₂, R³, or CONR′₂; O—(C₁-C₃)-alkyl; O—(C₁-C₃)-alkyloptionally substituted with NR′₂, OR′, CO₂R′, S (O₂)N(R′)₂, N═C—N(R′)₂,R³, or CONR′₂; NR′₂; OCF₃; CF₃; NO₂; CO₂R′; CONR′; R³; OR³; NR³; SR³;C(O)R³; C(O)N(R′)R³; C(O)OR³; SR′; S(O₂)N(R′)₂; SCF₃; N═C—N(R′)₂; or CN.

Q₂′ is selected from phenyl or a 5-6 member aromatic heterocyclic ringoptionally substituted with 1-3 substituents, each of which isindependently selected from halogen; C₁-C₃ alkyl optionally substitutedwith NR′₂, OR′, CO₂R′, CONR′₂, or O—P(O₃)H₂; O—(C₂-C₃)-alkyl optionallysubstituted with NR′₂, OR′, CO₂R′, CONR′₂, or OP(O₃)H₂; OCF₃; CF₃; OR⁴;O—CO₂R⁴; O—P(O₃)H₂; CO₂R′; CONR′; SR′; S(O₂)N(R′)₂; SCF₃; CN;N(R′)C(O)R⁴; N(R′)C(O)OR⁴; N(R′)C(O)C(O)R⁴; N(R′)S(O₂)R⁴; N(R′)R⁴;N(R⁴)₂; OR⁴; OC(O)R⁴; OP(O)₃H₂; or N═C—N(R′)₂; provided that Q₂′ is notphenyl optionally substituted 1 to 3 substituents independently selectedfrom halo, methoxy, cyano, nitro, amino, hydroxy, methyl or ethyl.

R′ is selected from hydrogen; (C₁-C₃)-alkyl; (C₂-C₃)-alkenyl or alkynyl;phenyl or phenyl substituted with 1 to 3 substituents independentlyselected from halo, methoxy, cyano, nitro, amino, hydroxy, methyl orethyl; or a 5-6 membered heterocyclic ring system optionally substitutedwith 1 to 3 substituents independently selected from halo, methoxy,cyano, nitro, amino, hydroxy, methyl or ethyl.

R³ is selected from 5-8 membered aromatic or non-aromatic carbocyclic orheterocyclic ring systems each optionally substituted with R′, R⁴,—C(O)R′, —C′(O)R⁴, —C(O)OR⁴ or -J; or an 8-10 membered bicyclic ringsystem comprising aromatic carbocyclic rings, aromatic heterocyclicrings or a combination of an aromatic carbocyclic ring and an aromaticheterocyclic ring each optionally substituted with R′, R⁴, —C(O)R′,—C(O)R⁴, —C(O)OR⁴ or -J.

R⁴ is (C₁-C₄)-straight or branched alkyl optionally substituted withN(R′)₂, OR′, CO₂R′, CON(R′)₂, or SO₂N(R²)₂; or a 5-6 memberedcarbocyclic or heterocyclic ring system optionally substituted withN(R′)₂, OR′, CO₂R′, CON(R′)₂, or SO₂N(R²)₂.

R⁵ is selected from hydrogen; (C₁-C₃)-alkyl optionally substituted withR³; (C₂-C₃)-alkenyl or alkynyl each optionally substituted with R³;phenyl or phenyl substituted with 1 to 3 substituents independentlyselected from halo, methoxy, cyano, nitro, amino, hydroxy, methyl orethyl; or a 5-6 membered heterocyclic ring system optionally substitutedwith 1 to 3 substituents independently selected from halo, methoxy,cyano, nitro, amino, hydroxy, methyl or ethyl.

W is selected from N(R²)SO₂—N(R²⁾ ₂; N(R²)SO₂—N(R²)(R³); N(R²)C(O)—OR²;N(R²)C(O)—N(R²)₂; N(R²)C(O)—N(R²)(R³); N(R²)C(O)—R²; N(R²)₂; C(O)—R²;CH(OH)—R²; C(O)—N(R²)₂; C(O)—OR²; J; or (C₁-C₄) straight or branchedalkyl optionally substituted with N(R′)₂, OR′, CO₂R′, CON(R′)₂, R³,SO₂N(R²)₂, OC(O)R², OC(O)R′, OC(O)N(R²)₂, —N(R⁴)(R⁵), —C(O)N(R⁵)(R²),—C(O)R⁵, —N(R²)C(O)N(R²)(R⁵), —NC(O)OR⁵, —OC(O)N(R²)(R⁵), or -J; a 5-6membered carbocyclic or heterocyclic ring system optionally substitutedwith N(R′)₂, OR′, CO₂R′, CON(R′)₂, or SO₂N(R²)₂; or a 8-10 memberedcarbocyclic or heterocyclic ring system optionally substituted withN(R′)₂, OR′, CO₂R′, CON(R′)₂, or SO₂N(R²)₂; provided that W is not an R³substituted C₁ alkyl.

W′ is selected from N(R²)—SO₂-Q₂; N(R²)—CO₂-Q₂; N(R²)—C(O)-Q₂;N(R²)(Q₂); C(O)-Q₂; CO₂-Q₂; C(O)N(R²)(Q₂); C(R²)₂Q₂.

Each R is independently selected from hydrogen, —R², —N(R²)₂, —OR′, SR²,—C(O)—N(R²)₂, —S(O₂)—N(R²)₂, —C(O)—OR² or —C(O)R² wherein two adjacent Rare optionally bound to one another and, together with each Y to whichthey are respectively bound, form a 4-8 membered carbocyclic orheterocyclic ring.

R² is selected from hydrogen, (C₁-C₃)-alkyl, or (C₁-C₃)-alkenyl; eachoptionally substituted with —N(R′)₂, —OR′, SR′, —C(O)—N(R′)₂,—S(O₂)—N(R′)₂, —C(O)—OR′, —NSO₂R⁴, —NSO₂R³, —C(O)N(R′)(R³), —NC(O)R⁴,—N(R′)(R³), —N(R′)(R⁴), —C(O)R³, —C(O)N(R′)(R⁴), —N(R⁴)₂, —C(O)N═C(NH)₂or R³.

Y is N or C.

Z is CH, N, C(OCH₃), C(CH₃), C(NH₂), C(OH) or C(F).

U is selected from R or W.

V is selected from —C(O)NH₂, —P(O)(NH₂)₂, or —SO₂NH₂.

A,B, and C are independently selected from —O—, —CHR′—, —CHR⁴—, —NR′—,—NR⁴— or —S—.

J is a (C₁-C₄) straight chain or branched alkyl derivative substitutedwith 1-3 substituents selected from D, -T-C(O)R′, or —OPO₃H₂.

D is selected from the group

T is either O or NH.

G is either NH₂ or OH.

In another embodiment, the invention provides pharmaceuticalcompositions comprising the p38 inhibitors of this invention. Thesecompositions may be utilized in methods for treating or preventing avariety of disorders, such as cancer, inflammatory diseases, autoimmunediseases, destructive bone disorders, proliferative disorders,infectious diseases, viral diseases and neurodegenerative diseases.These compositions are also useful in methods for preventing cell deathand hyperplasia and therefore may be used to treat or preventreperfusion/ischemia in stroke, heart attacks, and organ hypoxia. Thecompositions are also useful in methods for preventing thrombin-inducedplatelet aggregation. Each of these above-described methods is also partof the present invention.

DETAILED DESCRIPTION OF THE INVENTION

These compounds have the general formula:

wherein each of Q₁ and Q₂ are independently selected from a phenyl or5-6 membered aromatic heterocyclic ring system, or a 8-10 memberedbicyclic ring system comprising aromatic carbocyclic rings, aromaticheterocyclic rings or a combination of an aromatic carbocyclic ring andan aromatic heterocyclic ring.

The rings that make up Q₁ are substituted with 1 to 4 substituents, eachof which is independently selected from halo; C₁-C₃ alkyl optionallysubstituted with NR′₂, OR′, CO₂R′ or CONR′₂; O—(C₁-C₃)-alkyl optionallysubstituted with NR′₂, OR′, CO₂R′ or CONR′₂; NR′₂; OCF₃; CF₃; NO₂;CO₂R′; CONR′; SR′; S(O₂)N(R′)₂; SCF₃; CN; N(R′)C(O)R⁴; N(R′)C(O)OR⁴;N(R′)C(O)C(O)R⁴; N(R′)S(O₂)R⁴; N(R′)R⁴; N(R⁴)₂; OR⁴; OC(O)R⁴; OP(O)₃H₂;or N═C—N(R′)₂.

The rings that make Up Q₂ are optionally substituted with up to 4substituents, each of which is independently selected from halo; C₁-C₃straight or branched alkyl optionally substituted with NR′₂, OR′, CO₂R′,S(O₂)N(R′)₂, N═C—N(R′)₂, R³, or CONR′₂; O—(C₁-C₃)-alkyl; O—(C₁-C₃)-alkyloptionally substituted with NR′₂, OR′, CO₂R′, S(O₂)N(R′)₂, N═C—N(R′)₂,R³, or CONR′₂; NR′₂; OCF₃; CF₃; NO₂; CO₂R′; CONR′; R³; OR³; NR³; SR³;C(O)R³; C(O)N(R′)R³; C(O)OR³; SR′; S(O₂)N(R′)₂; SCF₃; N═C—N(R′)₂; or CN.

Q₂′ is selected from phenyl or a 5-6 member aromatic heterocyclic ringoptionally substituted with 1-3 substituents, each of which isindependently selected from halogen; C₁-C₃ alkyl optionally substitutedwith NR′₂, OR′, CO₂R′, CONR′₂, or O—P(O₃)H₂; O—(C₂-C₃)-alkyl optionallysubstituted with NR′₂, OR′, CO₂R′, CONR′₂, or OP(O₃)H₂; OCF₃; CF₃; OR⁴;O—CO₂R⁴; O—P(O₃)H₂; CO₂R′; CONR′; SR′; S(O₂)N(R′)₂; SCF₃; CN;N(R′)C(O)R⁴; N(R′)C(O)OR⁴; N(R′)C(O)C(O)R⁴; N(R′)S(O₂)R⁴; N(R′)R⁴;N(R⁴)₂; OR⁴; OC(O)R⁴; OP(O)₃H₂; or N═C—N(R′)₂; provided that Q₂′ is notphenyl optionally substituted 1 to 3 substituents independently selectedfrom halo, methoxy, cyano, nitro, amino, hydroxy, methyl or ethyl.

R′ is selected from hydrogen; (C₁-C₃)-alkyl; (C₂-C₃)-alkenyl or alkynyl;phenyl or phenyl substituted with 1 to 3 substituents independentlyselected from halo, methoxy, cyano, nitro, amino, hydroxy, methyl orethyl; or a 5-6 membered heterocyclic ring system optionally substitutedwith 1 to 3 substituents independently selected from halo, methoxy,cyano, nitro, amino, hydroxy, methyl or ethyl.

R³ is selected from 5-8 membered aromatic or non-aromatic carbocyclic orheterocyclic ring systems each optionally substituted with R′, R⁴,—C(O)R′, —C(O)R⁴, —C(O)OR⁴ or -J; or an 8-10 membered bicyclic ringsystem comprising aromatic carbocyclic rings, aromatic heterocyclicrings or a combination of an aromatic carbocyclic ring and an aromaticheterocyclic ring each optionally substituted with R′, R⁴, —C(O)R′,—C(O)R⁴, —C(O)OR⁴ or -J.

R⁴ is (C₁-C₄)-straight or branched alkyl optionally substituted withN(R′)₂, OR′, CO₂R′, CON(R′)₂, or SO₂N(R²)₂; or a 5-6 memberedcarbocyclic or heterocyclic ring system optionally substituted withN(R′)₂, OR′, CO₂R′, CON(R′)₂, or SO₂N(R²)₂.

R⁵ is selected from hydrogen; (C₁-C₃)-alkyl optionally substituted withR³; (C₂-C₃)-alkenyl or alkynyl each optionally substituted with R³;phenyl or phenyl substituted with 1 to 3 substituents independentlyselected from halo, methoxy, cyano, nitro, amino, hydroxy, methyl orethyl; or a 5-6 membered heterocyclic ring system optionally substitutedwith 1 to 3 substituents independently selected from halo, methoxy,cyano, nitro, amino, hydroxy, methyl or ethyl.

W is selected from N(R²)SO₂—N(R²)₂; N(R²)SO₂—N(R²)(R³); N(R²)C(O)—OR²;N(R²)C(O)—N(R²)₂; N(R²)C(O)—N(R²)(R³); N(R²)C(O)—R²; N(R²)₂; C(O)—R²;CH(OH)—R²; C(O)—N(R²)₂; C(O)—OR²; J; or (C₁-C₄) straight or branchedalkyl optionally substituted with N(R′)₂, OR′, CO₂R′, CON(R′)₂, R³,SO₂N(R²)₂, OC(O)R², OC(O)R′, OC(O)N(R²)₂, —N(R⁴)(R⁵), —C(O)N(R⁵)(R²),—C(O)R⁵, —N(R²)C(O)N(R²)(R⁵), —NC(O)OR⁵, —OC(O)N(R²)(R⁵), or -J; a 5-6membered carbocyclic or heterocyclic ring system optionally substitutedwith N(R′)₂, OR′, CO₂R′, CON(R′)₂, or SO₂N(R²)₂; or a 8-10 memberedcarbocyclic or heterocyclic ring system optionally substituted withN(R′)₂, OR′, CO₂R′, CON(R′)₂, or SO₂N(R²)₂; provided that W is not an R³substituted C₁ alkyl.

W′ is selected from N(R²)—SO₂-Q₂; N(R²)—CO₂-Q₂; N(R²)—C(O)-Q₂;N(R²)(Q₂); C(O)-Q₂; CO₂-Q₂; C(O)N(R²)(Q₂); C(R² )₂Q₂.

Each R is independently selected from hydrogen, —R², —N(R²)₂, —OR², SR²,—C(O)—N(R²)₂, —S(O₂)—N(R²)₂, —C(O)—OR² or —C(O)R² wherein two adjacent Rare optionally bound to one another and, together with each Y to whichthey are respectively bound, form a 4-8 membered carbocyclic orheterocyclic ring.

When the two R components form a ring together with the Y components towhich they are respectively bound, it will obvious to those skilled inthe art that a terminal hydrogen from each unfused R component will belost. For example, if a ring structure is formed by binding those two Rcomponents together, one being —NH—CH₃ and the other being —CH₂—CH₃, oneterminal hydrogen on each R component (indicated in bold) will be lost.Therefore, the resulting portion of the ring structure will have theformula —NH—CH₂—CH₂—CH₂—.

R² is selected from hydrogen, (C₁-C₃)-alkyl, or (C₁-C₃)-alkenyl; eachoptionally substituted with —N(R′)₂, —OR′, SR′, —C(O)—N(R′)₂,—S(O₂)—N(R′)₂, —C(O)—OR′, —NSO₂R⁴, —NSO₂R³, —C(O)N(R′)(R³), —NC(O)R⁴,—N(R′)(R³), —N(R′)(R⁴), —C(O)R³, —C(O)N(R′)(R⁴) , —N(R⁴)₂, —C(O)N═C(NH)₂or R³.

Y is N or C.

Z is CH, N, C(OCH₃), C(CH₃), C(NH₂), C(OH) or C(F).

U is selected from R or W.

V is selected from —C(O)NH₂, —P(O)(NH₂)₂, or —SO₂NH₂.

A,B, and C are independently selected from —O—, —CHR′—, —CHR⁴—, —NR′—,—NR⁴— or —S—.

J is a (C₁-C₄) straight chain or branched alkyl derivative substitutedwith 1-3 substituents selected from D, -T-C(O)R′, or —OPO₃H₂.

D is selected from the group

T is either O or NH.

G is either NH₂ or OH.

According to a preferred embodiment, Q₁ is selected from phenyl orpyridyl containing 1 to 3 substituents, wherein at least one of saidsubstituents is in the ortho position and said substituents areindependently selected from chloro, fluoro, bromo, —CH₃, —OCH₃, —OH,—CF₃, —OCF₃, —O(CH₂)₂CH₃, NH₂, 3,4-methylenedioxy, —N(CH₃)₂,—NH—S(O)₂-phenyl, —NH—C(O)O—CH₂-4-pyridine, —NH—C(O)CH₂-morpholine,—NH—C(O)CH₂—N(CH₃)₂, —NH—C(O)CH₂-piperazine, —NH—C(O)CH₂-pyrrolidine,—NH—C(O)C(O)-morpholine, —NH—C(O)C(O)-piperazine,—NH—C(O)C(O)-pyrrolidine, —O—C(O)CH₂—N(CH₃)₂, or —O—(CH₂)₂—N(CH₃)₂.

Even more preferred are phenyl or pyridyl containing at least 2 of theabove-indicated substituents both being in the ortho position.

Some specific examples of preferred Q₁ are:

Most preferably, Q₁ is selected from 2-fluoro-6-trifluoromethylphenyl,2,6-difluorophenyl, 2,6-dichlorophenyl, 2-chloro-4-hydroxyphenyl,2-chloro-4-aminophenyl, 2,6-dichloro-4-aminophenyl,2,6-dichloro-3-aminophenyl, 2,6-dimethyl-4-hydroxyphenyl,2-methoxy-3,5-dichloro-4-pyridyl, 2-chloro-4,5 methylenedioxy phenyl, or2-chloro-4-(N-2-morpholino-acetamido)phenyl.

According to a preferred embodiment, Q₂ is phenyl, pyridyl or naphthylcontaining 0 to 3 substituents, wherein each substituent isindependently selected from chloro, fluoro, bromo, methyl, ethyl,isopropyl, —OCH₃, —OH, —NH₂, —CF₃, —OCF₃, —SCH₃, —OCH₃, —C(O)OH,—C(O)OCH₃, —CH₂NH₂, —N(CH₃)₂, —CH₂-pyrrolidine and —CH₂OH.

Some specific examples of preferred Q₂ are:

unsubstituted 2-pyridyl or unsubstituted phenyl.

Most preferred are compounds wherein Q₂ is selected from phenyl,2-isopropylphenyl, 3,4-dimethylphenyl, 2-ethylphenyl, 3-fluorophenyl,2-methylphenyl, 3-chloro-4-fluorophenyl, 3-chlorophenyl,2-carbomethoxylphenyl, 2-carboxyphenyl, 2-methyl-4-chlorophenyl,2-bromophenyl, 2-pyridyl, 2-methylenehydroxyphenyl, 4-fluorophenyl,2-methyl-4-fluorophenyl, 2-chloro-4-fluorphenyl, 2,4-difluorophenyl,2-hydroxy-4-fluorphenyl, 2-methylenehydroxy-4-fluorophenyl, 1-naphthyl,3-chloro-2-methylenehydroxy, 3-chloro-2-methyl, or 4-fluoro-2-methyl.

According to another preferred embodiment, each Y is C.

According an even more preferred embodiment, each Y is C and the R and Uattached to each Y component is selected from hydrogen or methyl.

According to another preferred embodiment, W is a 0-4 atom chainterminating in an alcohol, amine, carboxylic acid, ester, amide, orheterocycle.

Some specific examples of preferred W are:

Most preferably, W is selected from:

U has the same preferred and most preferred embodiments as W.

According to an even more preferred embodiment, each Y is C, and Wand/or U is not hydrogen.

Some preferred embodiments are provided in Table 1 to 6 below: TABLE 1Cmpd Number Structure VRT-042175

VRT-041238

VRT-042305

VRT-043675

VRT-042313

VRT-101257

VRT-101262

VRT-043176

VRT-043180

VRT-043181

VRT-101259

VRT-042196

TABLE 2 Cmpd Number Structure VRT-043188

VRT-100306

VRT-043190

VRT-043192

VRT-043672

VRT-043673

VRT-043674

VRT-100318

VRT-101256

VRT-101255

VRT-101253

VRT-101251

TABLE 3 Cmpd Number Structure VRT-100325

VRT-043683

VRT-101248

VRT-043685

VRT-043686

VRT-043690

VRT-100324

VRT-101249

VRT-043693

VRT-043694

VRT-043695

VRT-101247

TABLE 4 Cmpd Number Structure VRT-100310

VRT-043678

VRT-043191

VRT-100019

VRT-100020

VRT-043688

VRT-043675

VRT-042307

VRT-040569

VRT-100025

TABLE 5 Cmpd Number Structure VRT-100026

VRT-100304

VRT-100305

VRT-041291

VRT-032884

VRT-034465

VRT-100313

VRT-100315

VRT-100317

VRT-37742

VRT-100323

VRT-100146

TABLE 6 Cmpd Number Structure VRT-101094

VRT-043631

VRT-043008

VRT-042266

VRT-042169

VRT-043605

VRT-100075

VRT-100076

VRT-100077

Particularly preferred embodiments include:

wherein X is H,

Particularly preferred embodiments also include:

wherein X is NH₂ or N(CH₃)₂;

wherein X is OH, NH₂, or N(CH₃)₂.

Other particularly preferred embodiments include:

wherein X is OH, NH₂, N(CH₃)₂,

Other particularly preferred embodiments include:

Other particularly preferred embodiments include:

wherein X is

Most preferred embodiments include:

According to another embodiment, the present invention provides methodsof producing the above-identified inhibitors of p38 of the formulae(Ia), (Ib), (Ic), (Id) and (Ie). Representative synthesis schemes forformula (Ia) are depicted below.

Schemes 1-3 illustrate the preparation of compounds in which W is eitheran amino, carboxyl or an aldehyde function. In each case the particularmoiety may be modified through chemistry well known in the literature.For example the final amino compounds D and N (schemes 1 and 4respectively) may be acylated, sulfonylated or alkylated to preparecompounds within the scope of W. In all schemes, the L1 and L2 groups onthe initial materials are meant to represent leaving groups ortho to thenitrogen atom in a heterocyclic ring. For example, compound A may be2,6-dichloro-3 nitro pyridine.

In Scheme 1, W is selected from amino-derivatized compounds such asN(R²)SO₂—N(R²)₂; N(R²)SO₂—N(R²)(R³); N(R²)C(O)—OR²; N(R²)C(O)—N(R² )₂;N(R²)C(O)—N(R²)(R³); N(R²)C(O)—R²; or N(R²)₂.

In Scheme 1, the Q₂ ring is introduced utilizing one of many reactionsknow in the art which result in the production of biaryl compounds. Oneexample may be the reaction of an aryl lithium compound with thepyridine intermediate A. Alternatively, an arylmetalic compound such asan aryl stannane or an aryl boronic acid may be reacted with the arylhalide portion (intermediate A) in the presence of a Pd^(o) catalyst toform product B. In the next step, a Q1 substituted derivative such as aphenyl acetonitrile derivative may be treated with a base such as sodiumhydride, sodium amide, LDA, lithium hexamethyldisilazide or any numberof other non-nucleophilic bases to deprotonate the position alpha to thecyano group, which represents a masked amide moiety. This anion is thencontacted with intermediate B to form C. The nitrile or equivalent groupof intermediate C is then hydrolyzed to form the amide and the nitrogroup is subjected to reducing conditions to form the amine intermediateD. Intermediate D is then used to introduce various functionalitydefined by W through chemistry such as acylation, sulfonylation oralkylation reactions well known in the literature. Depending on theregiochemistry of the first two steps of this procedure, the first twosteps may need to be reversed.

In Scheme 2, W is selected from carboxyl-derivatized compounds such asC(O)—R²; CH(OH)—R²; C(O)—N(R²)₂; or C(O)—OR².

Scheme 2 generally follows the procedures described for Scheme 1 exceptthat a carboxyl intermediate such as E is the starting material. Thefirst two steps mirror Scheme 1, and, as mentioned for Scheme 1, may bereversed depending on the regiochemistry of specific examples.Intermediate G is formed from these first two steps and this materialmay be hydrolyzed as mention to for the carboxyl intermediate H. Thecarboxyl group may then be modified according to well-known proceduresfrom the literature to prepare analogs with defined W substituents suchas acylations, amidations and esterifications.

In Scheme 3, W is selected from (C₁-C₄) straight or branched alkyloptionally substituted with N(R′)₂, OR′, CO₂R′, CON(R′)₂, R³, orSO₂N(R²)₂; or a 5-6 membered carbocyclic or heterocyclic ring systemoptionally substituted with N(R′)₂, OR′, CO₂R′, CON(R′)₂, or SO₂N(R²)₂;provided that W is not an R³ substituted C₁ alkyl.

In scheme 3 a pyridine derivative is metalated and quenched with one ofmany known electrophiles which can generate an aldehyde, to formintermediate I. The aldehyde can then be masked to form the dimethylacetal J. This intermediate is then carried on as described in scheme 1and 2 to introduce the Q1 and Q2 substituents, to produce intermediateL. As before, these two steps may be interchanged depending on specificregiochemistry. The masked aldehyde of L may then be deprotected andutilized to form compounds with the defined W substitution using wellknow chemistry such as alkylations and reductive aminations.

Schemes 4-6 are similar to schemes 1-3 with the exception that thetargeted compounds are those in which Z=Nitrogen. The steps for theseschemes parallel 1-3 with the exception that the alkylation utilizing aphenyl acetonitrile is replaced with a reaction with a Q1 aminederivative such as a substituted aniline derivative. The amide portionof the molecule is then introduced in an acylation reaction with, forexample, chlorosulfonyl isocyanate.

In Scheme 4, W is selected from amino-derivatized groups such asN(R²)SO₂—N(R²)₂; N(R²)SO₂—N(R²)(R³); N(R²)C(O)—OR²; N(R²)C(O)—N(R²)₂;N(R²)C(O)—N(R²)(R³); N(R²)C(O)—R²; or N(R²)₂.

In Scheme 4, intermediate B (from scheme 1) is treated with, forexample, an aniline derivative in the presence of a base such aspotassium carbonate. Additionally, a palladium catalyst may be utilizedto enhance the reactivity of this general type of reaction, if needed.The resulting amine derivative is then acylated to form intermediate M.The nitro group of M is then reduced to form N and the amino group maythen be derivatized as described for scheme 1. As mentioned for schemes1-3, the steps involved in the introduction of the Q1 and Q2substituents may be interchanged depending on the specificregiochemistry of specific compounds.

In Scheme 5, W is selected from carboxyl-derivatized groups such asC(O)—R²; CH(OH)—R²; C(O)—N(R²)₂; or C(O)—OR².

In Scheme 6, W is selected from (C₁-C₄) straight or branched alkyloptionally substituted with N(R′)₂, OR′, CO₂R′, CON(R′)₂, R³, orSO₂N(R²) ₂; or a 5-6 membered carbocyclic or heterocyclic ring systemoptionally substituted with N(R′)₂, OR′, CO₂R′, CON(R′)₂, or SO₂N(R²)₂;provided that W is not an R³ substituted C₁ alkyl.

Schemes 5 and 6 generally follow the procedures mentioned above.

One having skill in the art will recognize schemes 1-6 may be used tosynthesize compounds having the general formula of (Ib), (Ic), (Id) and(Ie).

According to another embodiment of the invention, the activity of thep38 inhibitors of this invention may be assayed in vitro, in vivo or ina cell line. In vitro assays include assays that determine inhibition ofeither the kinase activity or ATPase activity of activated p38.Alternate in vitro assays quantitate the ability of the inhibitor tobind to p38 and may be measured either by radiolabelling the inhibitorprior to binding, isolating the inhibitor/p38 complex and determiningthe amount of radiolabel bound, or by running a competition experimentwhere new inhibitors are incubated with p38 bound to known radioligands.

Cell culture assays of the inhibitory effect of the compounds of thisinvention may determine the amounts of TNF, IL-1, IL-6 or IL-8 producedin whole blood or cell fractions thereof in cells treated with inhibitoras compared to cells treated with negative controls. Level of thesecytokines may be determined through the use of commercially availableELISAs.

An in vivo assay useful for determining the inhibitory activity of thep38 inhibitors of this invention are the suppression of hind paw edemain rats with Mycobacterium butyricum-induced adjuvant arthritis. This isdescribed in J. C. Boehm et al., J. Med. Chem., 39, pp. 3929-37 (1996),the disclosure of which is herein incorporated by reference. The p38inhibitors of this invention may also be assayed in animal models ofarthritis, bone resorption, endotoxin shock and immune function, asdescribed in A. M. Badger et al., J. Pharmacol.. ExperimentalTherapeutics, 279, pp. 1453-61 (1996), the disclosure of which is hereinincorporated by reference.

The p38 inhibitors or pharmaceutical salts thereof may be formulatedinto pharmaceutical compositions for administration to animals orhumans. These pharmaceutical compositions, which comprise an amount ofp38 inhibitor effective to treat or prevent a p38-mediated condition anda pharmaceutically acceptable carrier, are another embodiment of thepresent invention.

The term “p38-mediated condition”, as used herein means any disease orother deleterious condition in which p38 is known to play a role. Thisincludes conditions known to be caused by IL-1, TNF, IL-6 or IL-8overproduction. Such conditions include, without limitation,inflammatory diseases, autoimmune diseases, destructive bone disorders,proliferative disorders, infectious diseases, neurodegenerativediseases, allergies, reperfusion/ischemia in stroke, heart attacks,angiogenic disorders, organ hypoxia, vascular hyperplasia, cardiachypertrophy, thrombin-induced platelet aggregation, and conditionsassociated with prostaglandin endoperoxidase synthase-2.

Inflammatory diseases which may be treated or prevented by the compoundsof this invention include, but are not limited to, acute pancreatitis,chronic pancreatitis, asthma, allergies, and adult respiratory distresssyndrome.

Autoimmune diseases which may be treated or prevented by the compoundsof this invention include, but are not limited to, glomerulonephritis,rheumatoid arthritis, systemic lupus erythematosus, scleroderma, chronicthyroiditis, Graves' disease, autoimmune gastritis, diabetes, autoimmunehemolytic anemia, autoimmune neutropenia, thrombocytopenia, atopicdermatitis, chronic active hepatitis, myasthenia gravis, multiplesclerosis, inflammatory bowel disease, ulcerative colitis, Crohn'sdisease, psoriasis, or graft vs. host disease.

Destructive bone disorders which may be treated or prevented by thecompounds of this invention include, but are not limited to,osteoporosis, osteoarthritis and multiple myeloma-related bone disorder.

Proliferative diseases which may be treated or prevented by thecompounds of this invention include, but are not limited to, acutemyelogenous leukemia, chronic myelogenous leukemia, metastatic melanoma,Kaposi's sarcoma, and multiple myeloma.

Angiogenic disorders which may be treated or prevented by the compoundsof this invention include solid tumors, ocular neovasculization,infantile haemangiomas.

Infectious diseases which may be treated or prevented by the compoundsof this invention include, but are not limited to, sepsis, septic shock,and Shigellosis.

Viral diseases which may be treated or prevented by the compounds ofthis invention include, but are not limited to, acute hepatitisinfection (including hepatitis A, hepatitis B and hepatitis C), HIVinfection and CMV retinitis.

Neurodegenerative diseases which may be treated or prevented by thecompounds of this invention include, but are not limited to, Alzheimer'sdisease, Parkinson's disease, cerebral ischemias or neurodegenerativedisease caused by traumatic injury.

“p38-mediated conditions” also include ischemia/reperfusion in stroke,heart attacks, myocardial ischemia, organ hypoxia, vascular hyperplasia,cardiac hypertrophy, and thrombin-induced platelet aggregation.

In addition, p38 inhibitors of the instant invention are also capable ofinhibiting the expression of inducible pro-inflammatory proteins such asprostaglandin endoperoxide synthase-2 (PGHS-2), also referred to ascyclooxygenase-2 (COX-2). Therefore, other “p38-mediated conditions”which may be treated by the compounds of this invention include edema,analgesia, fever and pain, such as neuromuscular pain, headache, cancerpain, dental pain and arthritis pain.

The diseases that may be treated or prevented by the p38 inhibitors ofthis invention may also be conveniently grouped by the cytokine (IL-1,TNF, IL-6, IL-8) that is believed to be responsible for the disease.

Thus, an IL-1-mediated disease or condition includes rheumatoidarthritis, osteoarthritis, stroke, endotoxemia and/or toxic shocksyndrome, inflammatory reaction induced by endotoxin, inflammatory boweldisease, tuberculosis, atherosclerosis, muscle degeneration, cachexia,psoriatic arthritis, Reiter's syndrome, gout, traumatic arthritis,rubella arthritis, acute synovitis, diabetes, pancreatic β-cell diseaseand Alzheimer's disease.

TNF-mediated disease or condition includes, rheumatoid arthritis,rheumatoid spondylitis, osteoarthritis, gouty arthritis and otherarthritic conditions, sepsis, septic shock, endotoxic shock, gramnegative sepsis, toxic shock syndrome, adult respiratory distresssyndrome, cerebral malaria, chronic pulmonary inflammatory disease,silicosis, pulmonary sarcoisosis, bone resorption diseases, reperfusioninjury, graft vs. host reaction, allograft rejections, fever andmyalgias due to infection, cachexia secondary to infection, AIDS, ARC ormalignancy, keloid formation, scar tissue formation, Crohn's disease,ulcerative colitis or pyresis. TNF-mediated diseases also include viralinfections, such as HIV, CMV, influenza and herpes; and veterinary viralinfections, such as lentivirus infections, including, but not limited toequine infectious anemia virus, caprine arthritis virus, visna virus ormaedi virus; or retrovirus infections, including feline immunodeficiencyvirus, bovine immunodeficiency virus, or canine immunodeficiency virus.

IL-8 mediated disease or condition includes diseases characterized bymassive neutrophil infiltration, such as psoriasis, inflammatory boweldisease, asthma, cardiac and renal reperfusion injury, adult respiratorydistress syndrome, thrombosis and glomerulonephritis.

In addition, the compounds of this invention may be used topically totreat or prevent conditions caused or exacerbated by IL-1 or TNF. Suchconditions include inflamed joints, eczema, psoriasis, inflammatory skinconditions such as sunburn, inflammatory eye conditions such asconjunctivitis, pyresis, pain and other conditions associated withinflammation.

In addition to the compounds of this invention, pharmaceuticallyacceptable salts of the compounds of this invention may also be employedin compositions to treat or prevent the above-identified disorders.

Pharmaceutically acceptable salts of the compounds of this inventioninclude those derived from pharmaceutically acceptable inorganic andorganic acids and bases. Examples of suitable acid salts includeacetate, adipate, alginate, aspartate, benzoate, benzenesulfonate,bisulfate, butyrate, citrate, camphorate, camphorsulfonate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptanoate, glycerophosphate, glycolate,hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide,hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oxalate,palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, salicylate, succinate, sulfate, tartrate,thiocyanate, tosylate and undecanoate. Other acids, such as oxalic,while not in themselves pharmaceutically acceptable, may be employed inthe preparation of salts useful as intermediates in obtaining thecompounds of the invention and their pharmaceutically acceptable acidaddition salts. Salts derived from appropriate bases include alkalimetal (e.g., sodium and potassium), alkaline earth metal (e.g.,magnesium), ammonium and N—(C1-4 alkyl)4+ salts. This invention alsoenvisions the quaternization of any basic nitrogen-containing groups ofthe compounds disclosed herein. Water or oil-soluble or dispersibleproducts may be obtained by such quaternization.

Pharmaceutically acceptable carriers that may be used in thesepharmaceutical compositions include, but are not limited to, ionexchangers, alumina, aluminum stearate, lecithin, serum proteins, suchas human serum albumin, buffer substances such as phosphates, glycine,sorbic acid, potassium sorbate, partial glyceride mixtures of saturatedvegetable fatty acids, water, salts or electrolytes, such as protaminesulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,sodium chloride, zinc salts, colloidal silica, magnesium trisilicate,polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol,sodium carboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat.

The compositions of the present invention may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, intrasternal, intrathecal,intrahepatic, intralesional and intracranial injection or infusiontechniques. Preferably, the compositions are administered orally,intraperitoneally or intravenously.

Sterile injectable forms of the compositions of this invention may beaqueous or oleaginous suspension. These suspensions may be formulatedaccording to techniques known in the art using suitable dispersing orwetting agents and suspending agents. The sterile injectable preparationmay also be a sterile injectable solution or suspension in a non-toxicparenterally-acceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose, any bland fixed oilmay be employed including synthetic mono- or di-glycerides. Fatty acids,such as oleic acid and its glyceride derivatives are useful in thepreparation of injectables, as are natural pharmaceutically-acceptableoils, such as olive oil or castor oil, especially in theirpolyoxyethylated versions. These oil solutions or suspensions may alsocontain a long-chain alcohol diluent or dispersant, such ascarboxymethyl cellulose or similar dispersing agents which are commonlyused in the formulation of pharmaceutically acceptable dosage formsincluding emulsions and suspensions. Other commonly used surfactants,such as Tweens, Spans and other emulsifying agents or bioavailabilityenhancers which are commonly used in the manufacture of pharmaceuticallyacceptable solid, liquid, or other dosage forms may also be used for thepurposes of formulation.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers commonly used include lactose andcorn starch. Lubricating agents, such as magnesium stearate, are alsotypically added. For oral administration in a capsule form, usefuldiluents include lactose and dried cornstarch. When aqueous suspensionsare required for oral use, the active ingredient is combined withemulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Alternatively, the pharmaceutical compositions of this invention may beadministered in the form of suppositories for rectal administration.These can be prepared by mixing the agent with a suitable non-irritatingexcipient which is solid at room temperature but liquid at rectaltemperature and therefore will melt in the rectum to release the drug.Such materials include cocoa butter, beeswax and polyethylene glycols.

The pharmaceutical compositions of this invention may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, the pharmaceutical compositions may beformulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this invention include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, the pharmaceutical compositions can be formulatedin a suitable lotion or cream containing the active components suspendedor dissolved in one or more pharmaceutically acceptable carriers.Suitable carriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical compositions may be formulated asmicronized suspensions in isotonic, pH adjusted sterile saline, or,preferably, as solutions in isotonic, pH adjusted sterile saline, eitherwith or without a preservative such as benzylalkonium chloride.Alternatively, for ophthalmic uses, the pharmaceutical compositions maybe formulated in an ointment such as petrolatum.

The pharmaceutical compositions of this invention may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

The amount of p38 inhibitor that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated, the particular mode of administration. Preferably, thecompositions should be formulated so that a dosage of between 0.01-100mg/kg body weight/day of the inhibitor can be administered to a patientreceiving these compositions.

It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,rate of excretion, drug combination, and the judgment of the treatingphysician and the severity of the particular disease being treated. Theamount of inhibitor will also depend upon the particular compound in thecomposition.

According to another embodiment, the invention provides methods fortreating or preventing a p38-mediated condition comprising the step ofadministering to a patient one of the above-described pharmaceuticalcompositions. The term “patient”, as used herein, means an animal,preferably a human.

Preferably, that method is used to treat or prevent a condition selectedfrom inflammatory diseases, autoimmune diseases, destructive bonedisorders, proliferative disorders, infectious diseases, degenerativediseases, allergies, reperfusion/ischemia in stroke, heart attacks,angiogenic disorders, organ hypoxia, vascular hyperplasia, cardiachypertrophy, and thrombin-induced platelet aggregation.

According to another embodiment, the inhibitors of this invention areused to treat or prevent an IL-1, IL-6, IL-8 or TNF-mediated disease orcondition. Such conditions are described above.

Depending upon the particular p38-mediated condition to be treated orprevented, additional drugs, which are normally administered to treat orprevent that condition, may be administered together with the inhibitorsof this invention. For example, chemotherapeutic agents or otheranti-proliferative agents may be combined with the p38 inhibitors ofthis invention to treat proliferative diseases.

Those additional agents may be administered separately, as part of amultiple dosage regimen, from the p38 inhibitor-containing composition.Alternatively, those agents may be part of a single dosage form, mixedtogether with the p38 inhibitor in a single composition.

In order that the invention described herein may be more fullyunderstood, the following examples are set forth. It should beunderstood that these examples are for illustrative purposes only andare not to be construed as limiting this invention in any manner.

EXAMPLE 1 Synthesis of p38 Inhibitor Compound 6

To a solution of LDA (60 mmol, 40 mLs) at −78° C., was added dropwise asolution of 2,6-Dibromopyridine (40 mmol, 9.48 gms) in THF (30 mLs,dried). The mixture was stirred at −78° C. for 20 minutes. Ethyl formate(400 mmol, 32.3 mLs) was added and stirring was continued at −78° C. for2 hours. Saturated ammonium chloride (200 mLs) was added and the mixturewas warmed to room temperature. The reaction mixture was diluted withethyl acetate and the organic layer was washed with aqueous acid andbase. The organic layer was dried and evaporated in vacuo. The resultingmaterial was purified by flash chromatography on silica gel followed byeluting with 10% ethyl acetate in n-hexane to afford 1 (32 mmol, 8.41gms) as a white solid.

A solution of 1 (13.08 mmol, 3.1 gms) and concentrated sulfuric acid (1mL) in methanol (50 mL) was refluxed overnight. The reaction mixture wascooled, neutralized with aqueous base and extracted into ethyl acetate.Drying and evaporation of the organic layer afforded 2 (11.77 mmol, 3.63gms) as an oil.

To a solution of t-Butoxide (2.2 mmol, 2 mLs) was added dropwise asolution of 2,6-Dichloroaniline (1.0 mmol, 162 mgs) in THF (2 mL,dried). The mixture was stirred at room temperature for 20 minutes. Asolution of 2 (1.0 mmol, 309 mgs) in THF (5 mLs) was added and stirringwas continued for 3 hours. The reaction mixture was diluted with ethylacetate and the organic layer was washed with aqueous acid and base. Theorganic layer was dried and evaporated in vacuo. The resulting materialwas purified by flash chromatography on silica gel followed by elutingwith 5% acetone in n-hexane to afford 3 (0.33 mmol, 128 mgs) as anorange solid.

o-Tolylboronic acid (0.34 mmol, 46 mgs), and 3 (0.20 mmol, 80 mgs) weredissolved in a toluene/ethanol (5/1) mixture. Thallium carbonate (0.5,235 mgs) and tetrakis(triphenylphosphine)palladium (0) (10 mgs) wasadded to the solution and the slurry was allowed to reflux for 30minutes. The reaction mixture was diluted with ethyl acetate and theorganic layer was washed with aqueous acid and base. The organic layerwas dried and evaporated in vacuo. The resulting material was purifiedby flash chromatography on silica gel followed by eluting with 5%methanol in methylene chloride to afford 4 (0.17 mmol, 61 mgs) as awhite solid.

A solution of 4 (0.17 mmol, 61 mgs) and chlorosulfonyl isocyanate (1mmol, 141.5 mgs) in methylene chloride (5 mLs) was stirred at roomtemperature overnight. The reaction mixture was diluted with ethylacetate and the organic layer was washed with aqueous acid and base. Theorganic layer was dried and evaporated in vacuo. The resulting materialwas purified by flash chromatography on silica gel followed by elutingwith 5% acetone in n-hexane to afford 5 (0.12 mmol, 46 mgs) as a whitesolid.

Sodium borohydride (1.0 mmol, 39.8 mgs) was added to a solution of 5(0.12 mmol, 46 mgs) in methanol (10 mLs) and the solution was stirredfor 15 minutes. The reaction was quenched with water. The reactionmixture was then diluted with ethyl acetate and the organic layer waswashed with aqueous acid and base. The organic layer was dried andevaporated in vacuo. The resulting material was purified by flashchromatography to afford 6 (0.08 mmol, 36 mgs) as a white solid.

The spectral data for compound 6 was:

¹H NMR (500 MHz, CDCl₃) δ7.90 (d, 1H), 7.60 (d, 2H), 7.5-7.3 (m, 5H),6.30 (d, 2H), 4.5 (s, 2H), 2.3 (s, 2H).

Synthesis of p38 Inhibitor Compound 7

The amino-alcohol (500 mg, 1. 43 mmol), which was prepared in the samemanner as 4, was dissolved in dichloromethane. Triethylamine (433 mg,4.29 mmol) was added, followed by acetyl chloride (168 mg, 2.15 mmol).The mixture was stirred at room temperature for one hour, poured intowater, and extracted with dichloromethane. The organic extract wasevaporated in vacuo and the residue was dissolved in 10.0 mL of toluene.A 20% solution of phosgene in toluene (5.0 mL) was added and thesolution was refluxed for two hours. The solution was cooled and 5.0 mLof concentrated ammonium hydroxide was added, precipitating a whitesolid. The mixture was poured into water and extracted with toluene. Theorganic extract was dried (MgSO₄) and evaporated in vacuo to afford 205mg of the urea-acetate 7 as a white solid.

The spectral data for compound 7 was:

¹H NMR (500 MHz, CDCl₃) δ7.80 (d, 1H), 7.62-7.50 (m, 2H), 7.25-7.0 (m,5H), 6.59 (d, 1H), 5.1 (s, 2H), 2.12 (s, 3H). HRMS showed MH+ 434.2 asthe major peak.

Synthesis of p38 Inhibitor Compound 8

The urea-alcohol (548 mg, 1.4 mmol), which was prepared in the samemanner as 6, was dissolved in 5.0 mL of toluene. A 20% solution ofphosgene in toluene (5.0 mL) was added and the solution was refluxed fortwo hours. The solution was cooled and 5.0 mL of concentrated ammoniumhydroxide was added, precipitating a white solid. The mixture was pouredinto water and extracted with toluene. The organic extract was dried(MgSO₄) and evaporated in vacuo to afford 284 mg of the carbamate 8 as awhite solid.

The spectral data of compound 8 was:

¹H NMR (500 MHz, CDCl₃) δ7.77 (d, 1H), 7.55-7.45 (m, 2H), 7.15-6.95 (m,5H), 6.50 (d, 1H), 5.40 (br s, 2H.), 5.00 (s, 2H). HRMS showed MH+ 435.1as the major peak.

EXAMPLE 2 Synthesis of p38 Inhibitor Compound 16

One equivalent of 2,6-dichloropyridine-4-carboxylic acid was dissolvedin THF. The solution was cooled to 0° C. and one equivalent of boranedimethyl sulfide complex was added. The solution was stirred at roomtemperature for twelve hours. The mixture was poured into water andextracted with diethyl ether. The ether extract was dried, andevaporated in vacuo to afford 9 in 93% yield.

One equivalent of 9 was dissolved in methylene chloride. One equivalentof methyl chloromethyl ether was added, followed by the addition of oneequivalent of ethyl diisopropylamine. The reaction was stirred at roomtemperature for several hours, poured into water and extracted with awater-immiscible solvent. The extract was dried and evaporated in vacuoto afford 10 in 86% yield.

One equivalent of potassium t-butoxide was added to a solution of oneequivalent of 2,6-dichlorophenyl acetonitrile in THF at roomtemperature. The mixture was stirred at room temperature for thirtyminutes, and a solution of the dichloropyridine 10 in THF was added.After stirring for 1.5 hours, the mixture was poured into aqueousammonium chloride and extracted with ethyl acetate. The extract wasdried and evaporated in vacuo. The residue was purified by flashchromatography to afford 11 in 79% yield as a white powder.

The acetal 11 was mixed with concentrated hydrochloric acid and stirredfor several hours. The mixture was extracted with a water-immicibleorganic solvent. The extract was washed with saturated aqueous NaHCO₃,dried, and evaporated in vacuo to afford 12.

The nitrile 12 was mixed with concentrated sulfuric acid and heated to100° C. for several minutes. The mixture was cooled, poured onto ice,and filtered to afford 13.

One equivalent of the chloropyridine 13 was dissolved in1,2-dimethoxyethane. One equivalent of 3-chloro-2-methylphenylboronicacid was added. A solution of one equivalent of sodium carbonate inwater was added along with a catalytic amount of tetrakis(triphenylphosphine) palladium (0). The mixture was heated to 80° C. forseveral hours. The mixture was poured into water and extracted with awater-immiscible organic solvent. The extract was dried, evaporated invacuo and purified by flash chromatography to afford 14.

One equivalent of the alcohol 14 was dissolved in THF. The solution wascooled to 0° C. and one equivalent of methanesulfonyl chloride was addedfollowing by one equivalent of triethylamine. The solution was stirredfor several hours, poured into water, and extracted with awater-immiscible solvent. The extract was dried and evaporated in vacuoto afford the crude mesylate 15.

One equivalent of the methanesulfonyl ester 15 was dissolved in THF. Thesolution was cooled to 0° C. and one equivalent of N-ethyl piperazinewas added following by one equivalent of triethylamine. The solution wasstirred for several hours, poured into water, and extracted with awater-immiscible solvent. The extract was dried, evaporated, andpurified by flash chromatography to afford the pure amine 16.

The spectral data for compound 16 is:

¹H NMR (500 MHz, CDCl₃) δ 9.85 (br s, 1H), 7.47 (dd, 1H), 7.42 (d, 1H),7.27 (m, 5H), 6.75 (s, 1H), 5.95 (s, 1H), 5.7 (br s, 1H), 3.5 (ABq, 2H),2.5-2.3 (m, 10H), 2.3 (s, 3H), 1.2 (t, 3H).

EXAMPLE 2 Cloning of p38 Kinase in Insect Cells

Two splice variants of human p38 kinase, CSBP1 and CSBP2, have beenidentified. Specific oligonucleotide primers were used to amplify thecoding region of CSBP2 cDNA using a HeLa cell library (Stratagene) as atemplate. The polymerase chain reaction product was cloned into thepET-15b vector (Novagen). The baculovirus transfer vector,pVL-(His)6-p38 was constructed by subcloning a XbaI-BamHI fragment ofpET15b-(His)6-p38 into the complementary sites in plasmid pVL1392(Pharmingen).

The plasmid pVL-(His)6-p38 directed the synthesis of a recombinantprotein consisting of a 23-residue peptide (MGSSHHHHHHSSGLVPRGSHMLE,where LVPRGS represents a thrombin cleavage site) fused in frame to theN-terminus of p38, as confirmed by DNA sequencing and by N-terminalsequencing of the expressed protein. Monolayer culture of Spodopterafrugiperda (Sf9) insect cells (ATCC) was maintained in TNM-FH medium(Gibco BRL) supplemented with 10% fetal bovine serum in a T-flask at 27°C. Sf9 cells in log phase were co-transfected with linear viral DNA ofAutographa califonica nuclear polyhedrosis virus (Pharmingen) andtransfer vector pVL-(His)6-p38 using Lipofectin (Invitrogen). Theindividual recombinant baculovirus clones were purified by plaque assayusing 1% low melting agarose.

EXAMPLE 3 Expression and Purification of Recombinant p38 Kinase

Trichoplusia ni (Tn-368) High-Five™ cells (Invitrogen) were grown insuspension in Excel-405 protein free medium (JRH Bioscience) in a shakerflask at 27° C. Cells at a density of 1.5×10⁶ cells/ml were infectedwith the recombinant baculovirus described above at a multiplicity ofinfection of 5. The expression level of recombinant p38 was monitored byimmunoblotting using a rabbit anti-p38 antibody (Santa CruzBiotechnology). The cell mass was harvested 72 hours after infectionwhen the expression level of p38 reached its maximum.

Frozen cell paste from cells expressing the (His)₆-tagged p38 was thawedin 5 volumes of Buffer A (50 mM NaH₂PO₄ pH 8.0, 200 mM NaCl, 2 mMβ-Mercaptoethanol, 10% Glycerol and 0.2 mM PMSF). After mechanicaldisruption of the cells in a microfluidizer, the lysate was centrifugedat 30,000×g for 30 minutes. The supernatant was incubated batchwise for3-5 hours at 4° C. with Talon™ (Clontech) metal affinity resin at aratio of 1 ml of resin per 2-4 mgs of expected p38. The resin wassettled by centrifugation at 500×g for 5 minutes and gently washedbatchwise with Buffer A. The resin was slurried and poured into a column(approx. 2.6×5.0 cm) and washed with Buffer A+5 mM imidazole.

The (His)₆-p38 was eluted with Buffer A+100 mM imidazole andsubsequently dialyzed overnight at 4° C. against 2 liters of Buffer B,(50 mM HEPES, pH 7.5, 25 mM β-glycerophosphate, 5% glycerol, 2 mM DTT).The His₆ tag was removed by addition of at 1.5 units thrombin(Calbiochem) per mg of p38 and incubation at 20° C. for 2-3 hours. Thethrombin was quenched by addition of 0.2 mM PMSF and then the entiresample was loaded onto a 2 ml benzamidine agarose (AmericanInternational Chemical) column.

The flow through fraction was directly loaded onto a 2.6×5.0 cmQ-Sepharose (Pharmacia) column previously equilibrated in Buffer B+0.2mM PMSF. The p38 was eluted with a 20 column volume linear gradient to0.6M NaCl in Buffer B. The eluted protein peak was pooled and dialyzedovernight at 4° C. vs. Buffer C (50 mM HEPES pH 7.5, 5% glycerol, 50 mMNaCl, 2 mM DTT, 0.2 mM PMSF).

The dialyzed protein was concentrated in a Centriprep (Amicon) to 3-4 mland applied to a 2.6×100 cm Sephacryl S-100HR (Pharmacia) column. Theprotein was eluted at a flow rate of 35 ml/hr. The main peak was pooled,adjusted to 20 mM DTT, concentrated to 10-80 mgs/ml and frozen inaliquots at −70° C. or used immediately.

EXAMPLE 4 Activation of p38

p38 was activated by combining 0.5 mg/ml p38 with 0.005 mg/ml DD-doublemutant MKK6 in Buffer B+10 mM MgCl₂, 2 mM ATP, 0.2 mM Na₂VO₄ for 30minutes at 20° C. The activation mixture was then loaded onto a 1.0×10cm MonoQ column (Pharmacia) and eluted with a linear 20 column volumegradient to 1.0 M NaCl in Buffer B. The activated p38 eluted after theADP and ATP. The activated p38 peak was pooled and dialyzed againstbuffer B+0.2 mM Na₂VO₄ to remove the NaCl. The dialyzed protein wasadjusted to 1.1M potassium phosphate by addition of a 4.0M stocksolution and loaded onto a 1.0×10 cm HIC (Rainin Hydropore) columnpreviously equilibrated in Buffer D (10% glycerol, 20 mMB-glycerophosphate, 2.0 mM DTT)+1.1MK₂HPO₄. The protein was eluted witha 20 column volume linear gradient to Buffer D+50 mM K₂HPO₄. The doublephosphorylated p38 eluted as the main peak and was pooled for dialysisagainst Buffer B+0.2 mM Na₂VO₄. The activated p38 was stored at −70° C.

EXAMPLE 5 p38 Inhibition Assays

A. Inhibition of Phosphorylation of EGF Receptor Peptide

This assay was carried out in the presence of 10 mM MgCl₂, 25 mMJ-glycerophosphate, 10% glycerol and 100 mM HEPES buffer at pH 7.6. Fora typical IC₅₀ determination, a stock solution was prepared containingall of the above components and activated p38 (5 nM). The stock solutionwas aliquotted into vials. A fixed volume of DMSO or inhibitor in DMSO(final concentration of DMSO in reaction was 5%) was introduced to eachvial, mixed and incubated for 15 minutes at room temperature. EGFreceptor peptide, KRELVEPLTPSGEAPNQALLR, a phosphoryl acceptor inp38-catalyzed kinase reaction (1), was added to each vial to a finalconcentration of 200 μM. The kinase reaction was initiated with ATP (100μM) and the vials were incubated at 30° C. After 30 minutes, thereactions were quenched with equal volume of 10% trifluoroacetic acid(TFA).

The phosphorylated peptide was quantified by HPLC analysis. Separationof phosphorylated peptide from the unphosphorylated peptide was achievedon a reverse phase column (Deltapak, 5 μm, C18 100D, Part no. 011795)with a binary gradient of water and acteonitrile, each containing 0.1%TFA. IC₅₀ (concentration of inhibitor yielding 50% inhibition) wasdetermined by plotting the percent (%) activity remaining againstinhibitor concentration.

B. Inhibition of ATPase Activity

This assay is carried out in the presence of 10 MM MgCl₂, 25 mMβ-glycerophosphate, 10% glycerol and 100 mM HEPES buffer at pH 7.6. Fora typical Ki determination, the Km for ATP in the ATPase activity ofactivated p38 reaction is determined in the absence of inhibitor and inthe presence of two concentrations of inhibitor. A stock solution isprepared containing all of the above components and activated p38 (60nM). The stock solution is aliquotted into vials. A fixed volume of DMSOor inhibitor in DMSO (final concentration of DMSO in reaction was 2.5%)is introduced to each vial, mixed and incubated for 15 minutes at roomtemperature. The reaction is initiated by adding various concentrationsof ATP and then incubated at 30° C. After 30 minutes, the reactions arequenched with 50 μl of EDTA (0.1 M, final concentration), pH 8.0. Theproduct of p38 ATPase activity, ADP, is quantified by HPLC analysis.

Separation of ADP from ATP is achieved on a reversed phase column(Supelcosil, LC-18, 3 μm, part no. 5-8985) using a binary solventgradient of following composition: Solvent A−0.1 M phosphate buffercontaining 8 mM tetrabutylammonium hydrogen sulfate (Sigma Chemical Co.,catalogue no. T-7158), Solvent B−Solvent A with 30% methanol.

Ki is determined from the rate data as a function of inhibitor and ATPconcentrations.

p38 inhibitors of this invention will inhibit the ATPase activity ofp38.

C. Inhibition of IL-1, TNF, IL-6 and IL-8 Production in LPS-StimulatedPBMCs

Inhibitors were serially diluted in DMSO from a 20 mM stock. At least 6serial dilutions were prepared. Then 4× inhibitor stocks were preparedby adding 4 μl of an inhibitor dilution to 1 ml of RPMI1640 medium/10%fetal bovine serum. The 4× inhibitor stocks contained inhibitor atconcentrations of 80 μM, 32 μM, 12.8 μM, 5.12 μM, 2.048 μM, 0.819 μM,0.328 μM, 0.131 μM, 0.052 μM, 0.021 μM etc. The 4× inhibitor stocks werepre-warmed at 37° C. until use.

Fresh human blood buffy cells were separated from other cells in aVacutainer CPT from Becton & Dickinson (containing 4 ml blood and enoughDPBS without Mg²⁺/Ca²⁺ to fill the tube) by centrifugation at 1500×g for15 min. Peripheral blood mononuclear cells (PBMCs), located on top ofthe gradient in the Vacutainer, were removed and washed twice withRPMI1640 medium/10% fetal bovine serum. PBMCs were collected bycentrifugation at 500×g for 10 min. The total cell number was determinedusing a Neubauer Cell Chamber and the cells were adjusted to aconcentration of 4.8×10⁶ cells/ml in cell culture medium (RPMI1640supplemented with 10% fetal bovine serum).

Alternatively, whole blood containing an anti-coagulant was useddirectly in the assay.

100 μl of cell suspension or whole blood were placed in each well of a96-well cell culture plate. Then 50 μl of the 4× inhibitor stock wasadded to the cells. Finally, 50 μl of a lipopolysaccharide (LPS) workingstock solution (16 ng/ml in cell culture medium) was added to give afinal concentration of 4 ng/ml LPS in the assay. The total assay volumeof the vehicle control was also adjusted to 200 μl by adding 50 μl cellculture medium. The PBMC cells or whole blood were then incubatedovernight (for 12-15 hours) at 37° C./5% CO₂ in a humidified atmosphere.

The next day the cells were mixed on a shaker for 3-5 minutes beforecentrifugation at 500×g for 5 minutes. Cell culture supernatants wereharvested and analyzed by ELISA for levels of IL-1b (R & D Systems,Quantikine kits, #DBL50), TNF-α (BioSource, #KHC3012), IL-6 (Endogen,#EH2-IL6) and IL-8 (Endogen, #EH2-IL8) according to the instructions ofthe manufacturer. The ELISA data were used to generate dose-responsecurves from which IC50 values were derived.

Results for the kinase assay (“kinase”;

subsection A, above), IL-1 and TNF in LPS-stimulated PBMCs (“cell”) andIL-1, TNF and IL-6 in whole blood (“WB”) for various p38 inhibitors ofthis invention are shown in Table 7 below: TABLE 7 Kinase Cell IL-1 CellTNF WB IL-1 WB TNF WB IL-6 Compound M.W. IC50 (uM) IC50 (uM) IC50 (uM)IC50 (uM) IC50 (uM) IC50 (uM) 17 402.28 0.056 0.021 0.14 0.42 0.064 0.2518 436.32 0.002 0.02 0.05 0.118 0.055 0.18 19 387.36 0.027 0.027 0.010.057 0.09 0.075

Other p38 inhibitors of this invention will also inhibit phosphorylationof EGF receptor peptide, and will inhibit the production of IL-1, TNFand IL-6, as well as IL-8, in LPS-stimulated PBMCs or in whole blood.

D. Inhibition of IL-6 and IL-8 Production in IL-1-Stimulated PBMCs

This assay is carried out on PBMCs exactly the same as above except that50 μl of an IL-1b working stock solution (2 ng/ml in cell culturemedium) is added to the assay instead of the (LPS) working stocksolution.

Cell culture supernatants are harvested as described above and analyzedby ELISA for levels of IL-6 (Endogen, #EH2-IL6) and IL-8 (Endogen,#EH2-IL8) according to the instructions of the manufacturer. The ELISAdata are used to generate dose-response curves from which IC50 valueswere derived.

E. Inhibition of LPS-Induced Prostaglandin Endoperoxide Synthase-2(PGHS-2, or COX-2) induction in PBMCs

Human peripheral mononuclear cells (PBMCs) are isolated from fresh humanblood buffy coats by centrifugation in a Vacutainer CPT (Becton &Dickinson). 15×10⁶ cells are seeded in a 6-well tissue culture dishcontaining RPMI 1640 supplemented with 10% fetal bovine serum, 50 U/mlpenicillin, 50 μg/ml streptomycin, and 2 mM L-glutamine. Compounds areadded at 0.2, 2.0 and 20 μM final concentrations in DMSO. LPS is thenadded at a final concentration of 4 ng/ml to induce enzyme expression.The final culture volume is 10 ml/well.

After overnight incubation at 37° C., 5% CO₂, the cells are harvested byscraping and subsequent centrifugation, the supernatant is removed, andthe cells are washed twice in ice-cold DPBS (Dulbecco's phosphatebuffered saline, BioWhittaker). The cells are lysed on ice for 10 min in50 μl cold lysis buffer (20 mM Tris-HCl, pH 7.2, 150 mM NaCl, 1%Triton-X-100, 1% deoxycholic acid, 0.1% SDS, 1 mM EDTA, 2% aprotinin(Sigma), 10 pg/ml pepstatin, 10 μg/ml leupeptin, 2 mM PMSF, 1 mMbenzamidine, 1 mM DTT) containing 1 μl Benzonase (DNAse from Merck). Theprotein concentration of each sample is determined using the BCA assay(Pierce) and bovine serum albumin as a standard. Then the proteinconcentration of each sample is adjusted to 1 mg/ml with cold lysisbuffer. To 100 μl lysate an equal volume of 2× SDS PAGE loading bufferis added and the sample is boiled for 5 min. Proteins (30 pg/lane) aresize-fractionated on 4-20% SDS PAGE gradient gels (Novex) andsubsequently transferred onto nitrocellulose membrane by electrophoreticmeans for 2 hours at 100 mA in Towbin transfer buffer (25 mM Tris, 192mM glycine) containing 20% methanol. After transfer, the membrane ispretreated for 1 hour at room temperature with blocking buffer (5%non-fat dry milk in DPBS supplemented with 0.1% Tween-20) and washed 3times in DPBS/0.1% Tween-20. The membrane is incubated overnight at 4°C. with a 1:250 dilution of monoclonal anti-COX-2 antibody (TransductionLaboratories) in blocking buffer. After 3 washes in DPBS/0.1% Tween-20,the membrane is incubated with a 1:1000 dilution of horseradishperoxidase-conjugated sheep antiserum to mouse Ig (Amersham) in blockingbuffer for 1 h at room temperature. Then the membrane is washed again 3times in DPBS/0.1% Tween-20. An ECL detection system (SuperSignal™CL-HRP Substrate System, Pierce) is used to determine the levels ofexpression of COX-2.

While we have hereinbefore presented a number of embodiments of thisinvention, it is apparent that our basic construction can be altered toprovide other embodiments which utilize the methods of this invention.

1. A compound of the formula:

wherein each of Q₁ and Q₂ are independently selected from a phenyl or5-6 membered aromatic heterocyclic ring system, or a 8-10 memberedbicyclic ring system comprising aromatic carbocyclic rings, aromaticheterocyclic rings or a combination of an aromatic carbocyclic ring andan aromatic heterocyclic ring; the rings that make up Q₁ are substitutedwith 1 to 4 substituents, each of which is independently selected fromhalo; C₁-C₃ alkyl optionally substituted with NR′₂, OR′, CO₂R′ orCONR′₂; O—(C₁-C₃)-alkyl optionally substituted with NR′₂, OR′, CO₂R′ orCONR′₂; NR′₂; OCF₃; CF₃; NO₂; CO₂R′; CONHR′; SR′; S(O₂)N(R′)₂; SCF₃; CN;N(R′)C(O)R⁴; N(R′)C(O)OR⁴; N(R′)C(O)C(O)R⁴; N(R′)S(O₂)R⁴; N(R′)R⁴;N(R⁴)₂; OR⁴; OC(O)R⁴; OP(O)₃H₂; N═CH—N(R′)₂; 3,4-methylenedioxy;—NH—C(O)O—CH₂-4-pyridine; —NH—C(O)CH₂-morpholine;—NH—C(O)CH₂-piperazine; or —NH—C(O)CH₂-pyrrolidine; the rings that makeup Q₂ are optionally substituted with up to 4 substituents, each ofwhich is independently selected from halo; C₁-C₃ straight or branchedalkyl optionally substituted with NR′₂, OR′, CO₂R′, S(O₂)N(R′)₂,N═CH—N(R′)₂, R³, or CONR′₂; O—(C₁-C₃)-alkyl; O—(C₁-C₃)-alkyl optionallysubstituted with NR′₂, OR′, CO₂R′, S(O₂)N(R′)₂, N═CH—N(R′)₂, R³, orCONR′₂; NR′₂; OCF₃; CF₃; NO₂; CO₂R′; CONHR′; R³; OR³; NHR³; SR³; C(O)R³;C(O)N(R′)R³; C(O)OR³; SR′; S(O₂)N(R′)₂; SCF₃; N═CH—N(R′)₂; CN;—NH—C(═NH)—NH₂; or —CH₂—NH—C(═NH)—NH₂; R′ is selected from hydrogen;(C₁-C₃)-alkyl; (C₂-C₃)-alkenyl or alkynyl; phenyl or phenyl substitutedwith 1 to 3 substituents independently selected from halo, methoxy,cyano, nitro, amino, hydroxy, methyl or ethyl; or a 5-6 memberedheterocyclic ring system optionally substituted with 1 to 3 substituentsindependently selected from halo, methoxy, cyano, nitro, amino, hydroxy,methyl or ethyl; R³ is selected from 5-8 membered aromatic ornon-aromatic carbocyclic or heterocyclic ring systems each optionallysubstituted with R′, R⁴, —C(O)R′, —C(O)R⁴, —C(O)OR⁴ or -J; or an 8-10membered bicyclic ring system comprising aromatic carbocyclic rings,aromatic heterocyclic rings or a combination of an aromatic carbocyclicring and an aromatic heterocyclic ring each optionally substituted withR′, R⁴, —C(O)R′, —C(O)R⁴, —C(O)OR⁴or -J; R⁴ is (C₁-C₄)-straight orbranched alkyl optionally substituted with N(R′)₂, OR′, CO₂R′, CON(R′)₂,or SO₂N(R²)₂; or a 5-6 membered carbocyclic or heterocyclic ring systemoptionally substituted with N(R′)₂, OR′, CO₂R′, CON(R′)₂, or SO₂N(R²)₂;each R is independently selected from hydrogen, —R , —N(R²)₂, —OR², SR²,—C(O)—N(R²)₂, —S(O₂)—N(R²)₂, —C(O)—OR² or —C(O)R² wherein two adjacent Rare optionally bound to one another and, together with each Y to whichthey are respectively bound, form a 4-8 membered carbocyclic orheterocyclic ring; R² is selected from hydrogen, (C₁-C₃)-alkyl, or(C₂-C₃)-alkenyl; each optionally substituted with —N(R′)₂, —OR′, SR′,—C(O)—N(R′)₂, —S(O₂)—N(R′)₂, —C(O)—OR′, —NHSO₂R⁴, —NHSO₂R³,—C(O)N(R′)(R³), —NHC(O)R⁴, —N(R′)(R³), —N(R′)(R⁴), —C(O)R³,—C(O)N(R′)(R⁴), -N(R⁴)₂, —C(O)N═C(NH)₂ or R³; J is a (C₁-C₄) straightchain or branched alkyl derivative substituted with 1-3 substituentsselected from D, -T-C(O)R′, or —OPO₃H₂; D is selected from the group

T is either O or NH; and G is either NH₂ or OH.
 2. The compoundaccording to claim 1, wherein Q₁ is selected from phenyl or pyridylcontaining 1 to 3 substituents independently selected from chloro,fluoro, bromo, —CH₃, —OCH₃, —OH, —CF₃, —OCF₃, —O(CH₂)₂CH₃, NH₂,3,4-methylenedioxy, —N(CH₃)₂, —NH—S(O)₂-phenyl,—NH—C(O)O—CH₂-4-pyridine, —NH—C(O)CH₂-morpholine, —NH—C(O)CH₂—N(CH₃)₂,—NH—C(O)CH₂-piperazine, —NH—C(O)CH₂-pyrrolidine,—NH—C(O)C(O)-morpholine, —NH—C(O)C(O)-piperazine,—NH—C(O)C(O)-pyrrolidine, —O—C(O)CH₂—N(CH₃)₂, or —O—(CH₂)₂—N(CH₃)₂ andwherein at least one of said substituents is in the ortho position. 3.The compound according to claim 2, wherein Q₁ contains at least twosubstituents, both of which are in the ortho position.
 4. The compoundaccording to claim 2, wherein Q₁ is selected from:


5. The compound according to claim 4, wherein Q₁ is selected from2-fluoro-6-trifluoromethylphenyl, 2,6-difluorophenyl,2,6-dichlorophenyl, 2-chloro-4-hydroxyphenyl, 2-chloro-4-aminophenyl,2,6-dichloro-4-aminophenyl, 2,6-dichloro-3-aminophenyl,2,6-dimethyl-4-hydroxyphenyl, 2-methoxy-3,5-dichloro-4-pyridyl,2-chloro-4,5 methylenedioxy phenyl, or2-chloro-4-(N-2-morpholino-acetamido)phenyl.
 6. The compound accordingto claim 1, wherein Q₂ is selected from phenyl, pyridyl or naphthyl andwherein Q₂ optionally contains up to 3 substituents, each of which isindependently selected from chloro, fluoro, bromo, methyl, ethyl,isopropyl, —OCH₃, —OH, —NH₂, —CF₃, —OCF₃, —SCH₃, —C(O)OH, —C(O)OCH₃,—CH₂NH₂, —N(CH₃)₂, —CH₂-pyrrolidine, —CH₂OH, —CH₂—N(CH₃)₂,—CH₂-piperazine, —NH—C(═NH)—NH₂, —CH₂—NH—C(═NH)—NH₂, and—CH₂—NH-imidazoline.
 7. The compound according to claim 6, wherein Q₂ isselected from:

unsubstituted 2-pyridyl or unsubstituted phenyl.
 8. The compoundaccording to claim 7, wherein Q₂ is selected from phenyl,2-isopropylphenyl, 3,4-dimethylphenyl, 2-ethylphenyl, 3-fluorophenyl,2-methylphenyl, 3-chloro-4-fluorophenyl, 3-chlorophenyl,2-carbomethoxylphenyl, 2-carboxyphenyl, 2-methyl-4-chlorophenyl,2-bromophenyl, 2-pyridyl, 2-methylenehydroxyphenyl, 4-fluorophenyl,2-methyl-4-fluorophenyl, 2-chloro-4-fluorophenyl, 2,4-difluorophenyl,2-hydroxy-4-fluorophenyl or 2-methylenehydroxy-4-fluorophenyl,1-naphthyl, 3-chloro-2-methylenehydroxy, 3-chloro-2-methyl, or4-fluoro-2-methyl. 9-24. (canceled)
 25. A pharmaceutical compositioncomprising an amount of a compound according to claim 1 effective toinhibit p38, and a pharmaceutically acceptable carrier.
 26. A method oftreating or preventing inflammatory diseases, autoimmune diseases,destructive bone disorders, proliferative disorders, infectiousdiseases, viral diseases neurodegenerative diseases, allergies,reperfusion/ischemia in stroke, myocardial ischemia, renal ischemia,heart attacks, angiogenic disorders, organ hypoxia, vascularhyperplasia, cardiac hypertrophy, thrombin-induced platelet aggregationor conditions associated with prostaglandin endoperoxide synthase-2 in apatient, said method comprising administering to said patient acomposition according to claim
 25. 27. The method according to claim 26,wherein said method is used to treat or prevent an inflammatory diseaseselected from acute pancreatitis, chronic pancreatitis, asthma,allergies, or adult respiratory distress syndrome.
 28. The methodaccording to claim 26, wherein said method is used to treat or preventan autoimmune disease selected from glomerulonephritis, rheumatoidarthritis, systemic lupus erythematosus, scleroderma, chronicthyroiditis, Graves' disease, autoimmune gastritis, diabetes, autoimmunehemolytic anemia, autoimmune neutropenia, thrombocytopenia, atopicdermatitis, chronic active hepatitis, myasthenia gravis, multiplesclerosis, inflammatory bowel disease, ulcerative colitis, Crohn'sdisease, psoriasis, or graft vs. host disease.
 29. The method accordingto claim 26, wherein said method is used to treat or prevent adestructive bone disorder selected from osteoarthritis, osteoporosis ormultiple myeloma-related bone disorder.
 30. The method according toclaim 26, wherein said method is used to treat or prevent aproliferative disease selected from acute myelogenous leukemia, chronicmyelogenous leukemia, metastatic melanoma, Kaposi's sarcoma, or multiplemyeloma.
 31. The method according to claim 26, wherein said method isused to treat or prevent an infectious disease selected from sepsis,septic shock, or Shigellosis.
 32. The method according to claim 26,wherein said method is used to treat or prevent a viral disease selectedfrom acute hepatitis infection, HIV infection or CMV retinitis.
 33. Themethod according to claim 26, wherein said method is used to treat orprevent a neurodegenerative disease selected from Alzheimer's disease,Parkinson's disease, cerebral ischemia or neurodegenerative diseasecaused by traumatic injury.
 34. The method according to claim 26,wherein said method is used to treat or prevent ischemia/reperfusion instroke or myocardial ischemia, renal ischemia, heart attacks, organhypoxia or thrombin-induced platelet aggregation.
 35. The methodaccording to claim 26, wherein said method is used to treat or prevent acondition associated with prostaglandin endoperoxide synthase-2 selectedfrom edema, fever, analgesia or pain.
 36. The method according to claim35, wherein said pain is selected from neuromuscular pain, headache,cancer pain, dental pain or arthritis pain.
 37. The method according toclaim 26, wherein said method is used to treat or prevent an angiogenicdisorder selected from solid tumors, ocular neovasculization, orinfantile haemangiomas.